BRIEF REPORT Proteasome Inhibitors Synergize with Tumor Necrosis Factor-Related Apoptosis-Induced Ligand to Induce Anaplastic Thyroid Carcinoma Cell Death

Context: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive types of cancer characterized by complete refractoriness to multimodal treatment approaches. Therapeutic strategies based on the simultaneous use of proteasome inhibitors and death receptor ligands have been shown to induce apoptosis in several tumor types but have not yet been explored in ATC. Objective and Methods: The aim of this study was to investigate the ability of the proteasome inhibitor Bortezomib to induce apoptosis in ATC cell lines. Bortezomib was used as a single agent or in combination with TNF-related apoptosis-induced ligand (TRAIL), a member of the TNF family that selectively induces tumor cell apoptosis. The molecular effects of Bortezomib were investigated by analyzing the expression of key regulators of cell cycle and apoptosis and the activation of different apoptotic pathways. Results: Bortezomib induced apoptosis in ATC cells at doses achieved in the clinical setting, differently from conventional chemotherapeutic agents. Simultaneous treatment with low doses of Bortezomib and TRAIL had a synergistic effect in inducing massive ATC cell apoptosis. Bortezomib increased the expression of cytotoxic TRAIL receptors, p21 (WAF/CIP1) and proapoptotic second mitochondria-derived activator of caspases/direct inhibitor of apoptosis binding protein with low pI, and reduced the expression of antiapoptotic mediators such as cellular Fas-associated death domain-like IL-1␤ converting enzyme inhibitory protein, Bcl-2, Bcl-X L , and inhibitor of apoptosis-1, thus resulting in cell death induction through the mitochondrial apoptotic pathway. Conclusions: The combination of proteasome inhibitors and TRAIL synergizes to induce the destruction of chemoresistant neoplastic thyrocytes and could represent a promising therapeutic strategy for the treatment of anaplastic thyroid carcinoma. (J Clin Endocrinol Metab 92: 1938 -1942, 2007

BRIEF REPORT Proteasome Inhibitors Synergize with Tumor Necrosis Factor-Related Apoptosis-Induced Ligand to Induce Anaplastic Thyroid Carcinoma Cell Death

Context: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive types of cancer characterized by complete refractoriness to multimodal treatment approaches. Therapeutic strategies based on the simultaneous use of proteasome inhibitors and death receptor ligands have been shown to induce apoptosis in several tumor types but have not yet been explored in ATC. Objective and Methods: The aim of this study was to investigate the ability of the proteasome inhibitor Bortezomib to induce apoptosis in ATC cell lines. Bortezomib was used as a single agent or in combination with TNF-related apoptosis-induced ligand (TRAIL), a member of the TNF family that selectively induces tumor cell apoptosis. The molecular effects of Bortezomib were investigated by analyzing the expression of key regulators of cell cycle and apoptosis and the activation of different apoptotic pathways. Results: Bortezomib induced apoptosis in ATC cells at doses achieved in the clinical setting, differently from conventional chemotherapeutic agents. Simultaneous treatment with low doses of Bortezomib and TRAIL had a synergistic effect in inducing massive ATC cell apoptosis. Bortezomib increased the expression of cytotoxic TRAIL receptors, p21 (WAF/CIP1) and proapoptotic second mitochondria-derived activator of caspases/direct inhibitor of apoptosis binding protein with low pI, and reduced the expression of antiapoptotic mediators such as cellular Fas-associated death domain-like IL-1␤ converting enzyme inhibitory protein, Bcl-2, Bcl-X L , and inhibitor of apoptosis-1, thus resulting in cell death induction through the mitochondrial apoptotic pathway. Conclusions: The combination of proteasome inhibitors and TRAIL synergizes to induce the destruction of chemoresistant neoplastic thyrocytes and could represent a promising therapeutic strategy for the treatment of anaplastic thyroid carcinoma. (J Clin Endocrinol Metab 92: 1938 -1942, 2007

In vitro evaluation of fresh sperm quality in tomcats: a comparison of two collection techniques

The collection of semen from tomcats by urethral catheterization (CT) after medetomidine administration offers a novel and easy approach to obtain good quality sperm for in vitro fertilization. This study was designed to compare the sperm quality parameters and in vitro fertilizing capacity of CT spermatozoa with those of spermatozoa retrieved after epididymal slicing (EP). Semen was collected in seventeen adult cats by urethral catheterization, after which the cat was orchiectomized. Motility, morphology, plasma membrane integrity, acrosomal status, and in vitro fertilizing capacity of both fresh CT and EP samples were evaluated. The results showed that both total and progressive motility, as well as the percentage of normal spermatozoa, were higher for EP sperm than for CT sperm (P 0.05) between CTand EP sperm. Nevertheless, no difference (P > 0.05) in in vitro fertilizing capacity between spermatozoa collected by means of the two different methods was found. In conclusion, semen collection by means of urethral catheterization after medetomidine administration yields fertilization results similar to epididymal slicing, despite the fact that several sperm variables were different. Since this novel catheterization technique is repeatable, is easy to perform and facilitates semen preparation protocols, it may be preferable for routine IVF experiments with fresh spermatozoa

WNT16 variants influence site-specific bone mass determination and fracture risk in Maltese postmenopausal women

BACKGROUND/INTRODUCTION: Epidemiological studies and translational models have highlighted the importance of WNT16 as a key regulator of bone mineral density (BMD).PURPOSE: The study aimed to investigate the effect of two WNT16 variants, a frameshift rs55710688 (insCCCA) in the Kozak sequence, and a single nucleotide variant rs3801387 (A>G) located in the last intron of WNT16, with BMD and fracture risk in the Maltese postmenopausal women.METHODS: Genotyping was performed in 1,045 women from the Malta Osteoporotic Fracture Study using Competitive Allele Specific PCR (rs55710688) and TaqMan® fluorogenic 5’ nuclease allelic discrimination (rs3801387). Genotype-phenotype associations were analysed using the Mann-Whitney statistic whereas odds ratios (OR) with 95% confidence intervals [CI] were computed by logistic regression analysis adjusted for confounders.RESULTS: Genotyping of the WNT16 rs55710688 and rs3801387 was successful in 1,038 (CCCA=24%) and 1,027 (G=27%) samples respectively. Women with the homozygous reference genotype for both WNT16 variants had a lower lumbar spine (LS) T-score relative to women with the homozygous alternative genotype (rs55710688 p=0.035; rs3801387 p=0.031). Risk ratios revealed that homozygosity for the reference alleles was associated with osteoporosis at the LS (rs55710688 adjusted-OR: 2.44 [1.16-5.13]; rs3801387: 2.40 [1.18-4.89]), and all-type of low-trauma fracture risk which was not attenuated by BMD (rs55710688: 2.17 [1.11-4.27]; rs3801387: 1.90 [1.05-3.55]). WNT16 rs55710688 reference genotype also exhibited a deleterious effect on femoral neck BMD (3.10 [1.05-9.15]). Finally, the haplotype with the reference alleles for WNT16 rs55710688 and rs3801387 was associated with LS BMD (p=0.007) and fracture risk (p=0.021).CONCLUSION(S): Results indicate that the WNT16 rs55710688 and rs3801387 variants are possible genetic determinants of site-specific BMD and fracture risk in Malta, which is in line with other epidemiological studies. Our findings support the results of in vitro assays and in silico modelling demonstrating reduced translational efficiency in the presence of the reference alleles culminating in lower bone formation.peer-reviewe

Proteasome inhibitors synergize with tumor necrosis factor-related apoptosis-induced ligand to induce anaplastic thyroid carcinoma cell death

Context: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive types of cancer characterized by complete refractoriness to multimodal treatment approaches. Therapeutic strategies based on the simultaneous use of proteasome inhibitors and death receptor ligands have been shown to induce apoptosis in several tumor types but have not yet been explored in ATC. Objective and Methods: The aim of this study was to investigate the ability of the proteasome inhibitor Bortezomib to induce apoptosis in ATC cell lines. Bortezomib was used as a single agent or in combination with TNF-related apoptosis-induced ligand (TRAIL), a member of the TNF family that selectively induces tumor cell apoptosis. The molecular effects of Bortezomib were investigated by analyzing the expression of key regulators of cell cycle and apoptosis and the activation of different apoptotic pathways. Results: Bortezomib induced apoptosis in ATC cells at doses achieved in the clinical setting, differently from conventional chemotherapeutic agents. Simultaneous treatment with low doses of Bortezomib and TRAIL had a synergistic effect in inducing massive ATC cell apoptosis. Bortezomib increased the expression of cytotoxic TRAIL receptors, p21 (WAF/CIP1) and proapoptotic second mitochondria-derived activator of caspases/direct inhibitor of apoptosis binding protein with low pI, and reduced the expression of antiapoptotic mediators such as cellular Fas-associated death domain-like IL-1\uce\ub2 converting enzyme inhibitory protein, Bcl-2, Bcl-XL, and inhibitor of apoptosis-1, thus resulting in cell death induction through the mitochondrial apoptotic pathway. Conclusions: The combination of proteasome inhibitors and TRAIL synergizes to induce the destruction of chemoresistant neoplastic thyrocytes and could represent a promising therapeutic strategy for the treatment of anaplastic thyroid carcinoma. Copyright \uc2\ua9 2007 by The Endocrine Society